Isolation of islets from human pancreas tissue for the treatment of Type 1 diabetes is hampered by the scarcity of the colleganase used in the enzymatic dispersion process. Currently available commercial preparations of collagenase require extensive screening and testing for its usefulness. The underlying reason for lot-to- lot variability is not understood. This proposal presents a potential solution to these problems by providing a production technique which could reproducibly and reliably provide consistently pure enzyme with collagenase activity in unlimited quantities. To achieve this goal, the contractor plans to identify and clone the genes responsible for collagenase production from Clostridium histolyticum. Crude preparations of collagenase will be first purified to homogeneity, an anti-collagenase antibody raised, and then used as a probe to identify the Clostridium,histolyticum gene. The recombinant colleganase will be expressed in Escherichia coli host and studied as to the effectiveness in producing dispersed islets.